Bicycle derivatives, their preparation and use

ABSTRACT

The description relates to bicyclene derivatives of formula (I) in which A, B, D, E, G, K, L, R 1 , R 2 , Z 1  and Z 2  have the meanings given in the description, and their production. These compounds are suitable for treating diseases. ##STR1##

This application is a 371 of PCT/EP 94/03980, filed 30 Nov. 1994 whichclaims priority of German application P 43 41 6659 filed Dec. 7, 1993.

The present invention relates to novel bicycle derivatives, theirpreparation and use in therapy.

Endothelin is a peptide which is composed of 21 amino acids and issynthesized and released by the vascular endothelium. Endothelin existsin three isoforms, ET-1, ET-2 and ET-3. "Endothelin" or "ET" hereinaftermeans one or all isoforms of endothelin. Endothelin is a potentvasoconstrictor and has a powerful effect on vascular tone. It is knownthat this vasoconstriction is caused by the binding of endothelin to itsreceptor (Nature, 332, 411≧415, 1988; FEBS Letters, 231, 440-444, 1988and Biochem. Biophys. Res. Commun., 154, 868-875, 1988).

Elevated or abnormal release of endothelin causes persistentvasoconstriction in peripheral, renal and cerebral blood vessels, whichmay lead to disorders. It has been reported in the literature thatelevated endothelin levels have been found in the plasma of patientswith hypertension, acute myocardial infarct, pulmonary hypertension,Raynaud's syndrome, atherosclerosis and in the airways of asthmatics(Japan J. Hypertension, 12, 79 (1989), J. Vascular Med. Biology 2, 207(1990), J. Am. Med. Association 264, 2868 (1990)).

Accordingly, substances which specifically inhibit the binding ofendothelin to the receptor ought also to antagonize the variousabovementioned physiological effects of endothelin and thereforerepresent valuable drugs.

We have found that certain bicycle derivatives have goodendothelin-antagonistic activity.

The invention relates to bicycle derivatives of the formula I ##STR2##where 2 of the radicals A, B, D and E are CH groups and the 2 otherradicals are CH groups or nitrogen atoms,

Z¹ is hydrogen or halogen, C₁₋₆ -alkyl, or phenyl, phenyl-C₁₋₄ -alkyl,naphthyl or naphthyl-C₁₋₆ -alkyl which is unsubstituted or substitutedon the aromatic radical by C₁₋₆ -alkyl, C₁₋₆ -alkoxy, halogen, CF₃, NO₂or CN, or C₂₋₆ -alkenyl or C₂₋₆ -alkynyl, C₃₋₇ -cycloalkyl or one of thegroups --NHR⁴, --NR⁴ ₂, --OR⁴, --SO₂ NHR⁴, --SO₂ NR⁴ ₂, --COR⁴ or --CO₂R⁴ (with R⁴ meaning C₁₋₄ -alkyl, phenyl, phenyl-C₁₋₄ -alkyl, naphthyl ornaphthyl-C₁₋₄ -alkyl),

Z² has one of the meanings indicated for Z¹ but is not hydrogen, or

Z¹ and Z² together with B and D are also one of the radicals ##STR3##(where Z³ has one of the meanings indicated for Z¹, and M is a CH₂ or NHgroup),

G is a direct linkage or the group CH--K (with K meaning hydrogen, C₁₋₆-alkyl, or a phenyl, benzyl, naphthyl or naphthylmethyl group which isunsubstituted or substituted in the aryl moiety by C₁₋₄ -alkyl, C₁₋₄-alkoxy, halogen, CF₃, NO₂ or CN),

K is alkyl or alkenyl with, in each case, up to 6 C atoms or the group##STR4## L is an alkylene, alkenylene or alkynylene group with, in eachcase, up to 6 C atoms or one of the groups ##STR5## (with R³ meaninghydrogen, C₁₋₄ -alkyl, benzyl or naphthylmethyl), ##STR6## (with Qmeaning C₁₋₆ -alkyl, aryl or CH₂ --R⁷ where R⁷ is phenyl or hetaryl) or##STR7## R¹ is --CO₂ R⁴ (with R⁴ meaning hydrogen, C₁₋₄ -alkyl orbenzyl), --CONR⁴ ₂, --OR⁴, --SR⁴, --SO₃ R⁴, --PO₃ R⁴ ₂ or tetrazolyl,and

R² is ##STR8## (with R⁵ and R⁶ meaning hydrogen, C₁₋₄ -alkyl, --OR⁴ or--SR⁴) or hetaryl,

and, where appropriate, the salts thereof with physiologically toleratedacids.

Preferred compounds of the formula I are those where one or more of themoieties A, B, D, E, G, K, L, R¹, R², Z¹ and Z² have the followingmeanings:

A,B,D --CH--

E --CH--, --N--

G --CH₂ -- or a direct linkage between N and CO

K --CH₂ -- ##STR9## R¹ --COOH R² ##STR10## (R³ ═H, C₁₋₃ -alkyl, --CHO,--COO--C₁₋₃ -alkyl), ##STR11## Z¹ C₁₋₆ -alkyl, C₂₋₆ -alkenyl, C₂₋₆-alkynyl, --NH--CO--C₂₋₅ -alkyl

Z² hydrogen

Z¹ and Z² a radical ##STR12## which is linked to B and D and issubstituted by one of the radicals mentioned as preferred for Z¹.

The compounds of the formula I are obtained, if E is nitrogen, by thefollowing route:

Reaction of a substituted amino carboxylic derivative of the formula IIwith an amine of the formula III results firstly in IV. Ring closurewith an activated derivative of carbonic acid affords the compounds ofthe formula V. Alkylation of the amide nitrogen leads to the compoundsof the formula I. The protective groups which are required whereappropriate on the intermediates in the individual reaction steps areintroduced and eliminated again by conventional methods. ##STR13##

The compounds of the general formula I are obtained, if E is methine orsubstituted methine, by the following route:

The carboxylic acid derivatives of the formula IV are converted into theN-acyl derivative VII by reaction with an activated halo carboxylic acidderivative of the formula VI. The activated halo carboxylic acidderivatives which are preferably used are the corresponding acidhalides. Cyclization is then carried out to give VIII with eliminationof hydrogen halide. Substitution on the amide nitrogen leads to thecompounds of the formula I. The protective groups which are required onthe intermediates in the individual reaction steps are introduced andeliminated again by conventional methods. ##STR14##

The compounds of the present invention provide a novel therapeuticpotential for the treatment of hypertension, pulmonary hypertension,myocardial infarct, angina pectoris, acute kidney failure, renalinsufficiency, cerebral vasospasms, cerebral ischemia, subarachnoidhemorrhages, migrane, asthma, atherosclerosis, endotoxic shock,endotoxin-induced organ failure, intravascular coagulation, restenosisafter angioplasty and cyclosporin-induced kidney failure, andhypertension.

The good effect of the compounds can be shown in the followingexperiments:

Receptor Binding Studies

For binding studies, cloned human ET_(A) receptor-expressing CHO cellsand guinea pig cerebellar membranes with >60% ET_(B) compared withET_(A) receptors were used.

Membrane Preparation

The ET_(A) receptor-expressing CHO cells were grown in F₁₂ medium with10% fetal calf serum, 1% glutamine, 100 U/ml penicillin and 0.2%streptomycin (Gibco BRL, Gaithersburg, Md., USA). After 48 h, the cellswere washed with PBS and incubated with 0.05% trypsin-containing PBS for5 min. Neutralization was then carried out with F₁₂ medium, and thecells were collected by centrifugation at 300×g. For lysis of thesecells, the pellet was briefly washed with lysis buffer (5 mM Tris-HCl,pH 7.4 with 10% glycerol) and then incubated in a concentration of 10⁷cells/ml of lysis buffer at 4° C. for 30 min. The membranes werecentrifuged at 20,000×g for 10 min, and the pellet was stored in liquidnitrogen.

Guinea pig cerebella were homogenized in a Potter-Elvejhem homogenizerand obtained by differential centrifugation at 1,000×g for 10 min andrenewed centrifugation of the supernatant at 20,000×g for 10 min.

Binding Assays

For the ET_(A) and ET_(B) receptor binding assay, the membranes weresuspended in incubation buffer (50 mM Tris-HCl, pH 7.4 with 5 mM MnCl₂,40 μg/ml bacitracin and 0.2% BSA) at a concentration of 50 μg of proteinper assay mixture and incubated at 25° C. with 25 pM ¹²⁵ I!-ET₁ (ET_(A)receptor assay) or 25 pM ¹²⁵ I!-RZ₃ (ET_(B) receptor assay) in thepresence and absence of test substance. The nonspecific binding wasdetermined with 10⁻⁷ M ET₁. After 30 min, the free and the boundradioligand were separated by filtration through GF/B glass fiberfilters (Whatman, England) on a Skatron cell collector (Skatron, Lier,Norway), and the filters were washed with ice-cold Tris-HCl buffer, pH7.4 with 0.2% BSA. The radioactivity collected on the filters wasquantified using a Packard 2200 CA liquid scintillation counter.

The K_(i) values were determined by a non-linear regression analysiswith the LIGAND program.

Functional in vitro assay system to search for endothelin receptor(subtype A) antagonists.

This assay system is a functional, cell-based assay for endothelinreceptors. Certain cells show an increase in the intracellular calciumconcentration when they are stimulated with endothelin 1 (ET1). Thisincrease can be measured in intact cells which have been loaded withcalcium-sensitive dyes.

1-fibroblasts which had been isolated from rats and on which anendogenous endothelin receptor of subtype A had been detected wereloaded with the fluorescent dye Fura 2-an as follows: Aftertrypsinization, the cells were resuspended in buffer A (120 mM NaCl, 5mM KCl, 1.5 mM MgCl₂, 1 mM CaCl₂, 25 mM HEPES, 10 mM glucose, pH 7.4) toa density of 2×10⁶ /ml and incubated with Fura 2-an (2 μM), PluronicF-127 (0.04%) and DMSO (0.2%) in the dark at 37° C. for 30 min. Thecells were then washed twice with buffer A and resuspended at 2×10⁶ /ml.

The fluorescence signal from 2×10⁵ cells per ml with Ex/Em 380/510 wasrecorded continuously at 30° C. The test substances and then, afterincubation for 3 min, ET1 were added to the cells, and the maximumchange in fluorescence was determined. The response of the cells to ET1without previous addition of a test substance served as control and wasset equal to 100%.

Testing of ET Antagonists In Vivo

Male SD rats weighing 250-300 g were anaesthetized with amobarbital,artificially ventilated, vagotomized and pithed. The carotid artery andjugular vein were catheterized.

In control animals, intravenous administration of 1 μg/kg of ET1 leadsto a distinct rise in blood pressure, which persists for a lengthyperiod.

The test animals received i.v. injection (1 ml/kg) of the test compounds5 min before administration of ET1. To determine the ET-antagonisticproperties, the rise in blood pressure in the test animals was comparedwith that in the control animals.

Endothelin-1--Induced Sudden Death in Mice

The principle of the test comprises the inhibition of the sudden heartdeath caused by endothelin in mice, probably owing to constriction ofthe coronary vessels, by pretreatment with endothelin receptorantagonists. Intravenous injection of 10 nmol/kg endothelin in a volumeof 5 ml/kg of body weight results in death of the animals within a fewminutes.

The lethal endothelin-1 dose is checked in each case on a small group ofanimals. If the test substance is administered intravenously, the lethalendothelin-1 injection in the reference group usually takes place 5 minthereafter. With other modes of administration, the predosage times arelonger, where appropriate up to several hours.

The survival rate is recorded and effective doses which protect 50% ofthe animals against endothelin-induced heart death for 24 h or longer(ED 50) are determined.

Functional Vessel Test for Endothelin Receptor Antagonists

Segments of rabbit aorta under an initial tension of 2 g after arelaxation time of 1 h in Krebs-Henseleit solution at 37° C. and a pH offrom 7.3 to 7.4 are initially induced to contract with K⁺. After washingout, an endothelin dose-effect plot is constructed up to the maximum.

Potential endothelin antagonists are administered to other preparationsof the same vessel 15 min before starting the endothelin dose-effectplot. The effects of the endothelin are calculated as a % of the K⁺-induced contraction. Effective endothelin antagonists shift theendothelin dose-effect plot to the right.

The novel compounds may have acidic or basic groups and may thereforeexist in the form of salts.

Particularly suitable physiologically tolerated acids for salt formationare: hydrochloric acid, hydroiodic acid, sulfuric acid, phosphoric acid,acetic acid, citric acid, malonic acid, salicylic acid, maleic acid,fumaric acid, succinic acid, ascorbic acid, malic acid, methanesulfonicacid, lactic acid, gluconic acid, glucuronic acid, sulfamic acid,benzoic acid, tartaric acid.

Examples of suitable bases are alkali metal and alkaline earth metalhydroxides.

The compounds according to the invention can be administered orally orparenterally (subcutaneously, intravenously, intramuscularly,intraperitoneally) in a conventional way. Administration can also takeplace with vapors or sprays through the nasal pharyngeal space.

The dosage depends on the age, condition and weight of the patient andon the mode of administration. As a rule, the daily dose of activesubstance is about 0.5-50 mg/kg of body weight on oral administrationand about 0.1-10 mg/kg of body weight on parenteral administration.

The novel compounds can be used in conventional solid or liquidpharmaceutical forms, eg. as uncoated or (film-)coated tablets,capsules, powders, granules, suppositories, solutions, ointments, creamsor sprays. These are produced in a conventional manner. The activesubstances can in these cases be processed with conventionalpharmaceutical aids such as tablet binders, bulking agents,preservatives, tablet disintegrants, flow regulators, plasticizers,wetting agents, dispersants, emulsifiers, solvents, release-slowingagents, antioxidants and/or propellant gases (cf. H. Sucker et al.:Pharmazeutische Technologie, Thieme-Verlag, Stuttgart, 1991). Theadministration forms obtained in this way normally contain from 0.1 to90% by weight of active substance.

EXAMPLE 1

a) 5-Iodoisatoic anhydride

10 g (0.038 mol) of 5-iodoanthranilic acid were dissolved in 180 ml ofTHF, and 3.76 g (0.0127 mol) of bis(trichloromethyl) carbonate dissolvedin 20 ml of THF were added. The mixture was then stirred at roomtemperature for 1 h and at 50° C. for 2 h. After cooling, theprecipitate was filtered off with suction and dried. 7.8 g (71%) of5-iodoisatoic anhydride were obtained as white crystals.

b) 2-Amino-5-iodo-N-(2-methoxycarbonylethyl)benzamide

7.28 g (25.2 mmol) of 5-iodoisatoic anhydride and 3.52 g (25.2 mmol) ofβ-alanine methyl ester hydrochloride were introduced into 60 ml of DMF,and 6.12 g (60.5 mmol) of triethylamine were added dropwise. The mixturewas heated at 50° C. for 4 h, cooled and added to a mixture of 15 ml of2N NaOH with ice. The aqueous phase was extracted with ethyl acetate,and the organic phase was dried and concentrated under reduced pressure.6.6 g of a yellow oil were obtained and were purified by chromatographyon silica gel with ethyl acetate/n-heptane (1:1). 4.6 g (52%) of2-amino-5-iodo-N-(2-methoxycarbonylethyl)benzamide were obtained with anRF of 0.40 (ethyl acetate/n-heptane, 2:1).

c) 6-Iodo-3-(2-methoxycarbonylethyl)quinazoline-2,4-dione

4.6 g of 2-amino-5-iodo-N-(2-methoxycarbonylethyl)benzamide wereintroduced with 2.66 g (26.3 mmol) of triethylamine into 50 ml ofdichloromethane and, under nitrogen, 1.21 g (4.1 mmol) ofbis(trichloromethyl) carbonate dissolved in 15 ml of dichloromethanewere added dropwise. The mixture was refluxed for 2 h and, aftercooling, added to ice-water. The aqueous phase was extracted withdichloromethane, and the organic phase was dried and concentrated. 3.64g (74%) of 6-iodo-3-(2-methoxycarbonylethyl)quinazoline-2,4-dione R_(F)0.52 (ethyl acetate/n-heptane, 2:1)! were obtained as a yellowish solid.

d)6-Iodo-3-(2-methoxycarbonylethyl)-1-(N-t-butylcarbamoyl-3-indolylmethyl)quinazoline-2,4-dione

2.6 g (7 mmol) of 6-iodo-3-(2-methoxycarbonylethyl)quinazoline-2,4-dioneand 1.95 g (14.1 mmol) of potassium carbonate were suspended in acetone.To this mixture were added 3.05 g (9.8 mmol) ofN-t-butylcarbamoyl-3-indolylmethyl bromide dissolved in 20 ml ofacetone, and the mixture was stirred at room temperature for 5 h. Thesolvent was then stripped off under reduced pressure, and the residuewas taken up in 50 ml of phosphate buffer (pH 7) and 150 ml of ethylacetate. The product was extracted with ethyl acetate, dried andconcentrated under reduced pressure. 5.9 g of a dark oil were obtainedand were chromatographed on silica gel with ethyl acetate/n-heptane(ratio 1:4) as eluent. 1.9 g (45%) of6-iodo-3-(2-methoxycarbonylethyl)-1-(N-t-butylcarbamoyl-3-indolylmethyl)quinazoline-2,4-dione,R_(F) 0.18 (ethyl acetate/n-heptane, 1:4) were obtained.

e)6-(1-Pentenyl)-3-(2-methoxycarbonylethyl)-1-(N-t-butylcarbamoyl-3-indolylmethyl)quinazoline-2,4-dione

1.09 g (1.8 mmol) of6-iodo-3-(2-methoxycarbonylethyl)-1-(N-t-butylcarbamoyl-3-indolylmethyl)quinazoline-2,4-dioneand 0.75 g (5.4 mmol) of potassium carbonate were introduced in 15 ml ofDMF, and 0.63 g (9.0 mmol) of 1-pentene, 0.58 g (1.8 mmol) oftetrabutylammonium bromide and 10 mg of palladium acetate were added.The mixture was stirred at room temperature for 48 h and thenconcentrated under reduced pressure. The crude product waschromatographed on silica gel, eluent dichloromethane (2% methanol).0.69 g (70%) of product, R_(F) 0.12 (dichloromethane+2% methanol) wasobtained.

f)6-(1-Pentenyl)-3-(2-methoxycarbonylethyl)-1-(3-indolylmethyl)quinazoline-2,4-dione

0.53 g of the product obtained in e) was dissolved in 5 ml ofdichloromethane, 1.13 g (9.9 mol) of trifluoroacetic acid were added,and the mixture was stirred at room temperature for 16 h. It was thenwashed with water, dried and concentrated. 0.45 g of crude product wasobtained and was reacted further without purification.

g)6-(1-Pentenyl)-3-(2-carboxyethyl)-1-(3-indolylmethyl)quinazoline-2,4-dione

0.45 g of the crude product obtained in f) was dissolved in 10 ml ofTHF, and 0.03 mg (1.25 mmol) of lithium hydroxide dissolved in 2 ml ofwater was added, and the mixture was stirred at room temperature for 16h. The solvent was stripped off under reduced pressure, and the residuewas taken up in ethyl acetate and water; the aqueous phase was adjustedto pH 9 with ammonia solution and extracted with ethyl acetate. Theorganic phase was dried and concentrated. 0.37 g of crude product wasobtained. HPLC (reversed phase material, acetonitrile/water) resulted in0.12 g (0.28 mmol) of6-(1-pentenyl)-3-(2-carboxyethyl)-1-(3-indolylmethyl)quinazoline-2,4-dione.

The following were prepared as in Example 1:

6-E-(3-methyl-1-butenyl)!-3-(2-carboxyethyl)-1-(3-indolylmethyl)quinazoline-2,4-dione

6- E-(2-p-(1,1-dimethylethyl)phenyl)!ethenyl-3-(2-carboxyethyl)-1-(3-indolylmethyl)quinazoline-2,4-dione

6-E-(4-methyl-1-pentenyl)!-3-(2-carboxyethyl)-1-(3-indolylmethyl)quinazoline-2,4-dione

6-E-(2-cyclohexylethenyl)!-3-(2-carboxyethyl)-1-(3-indolylmethyl)quinazoline-2,4-dione,melting point 122°-126° C.

6-E-(1-pentenyl)!-3-(carboxymethyl)-1-(N-methyl-3-indolylmethyl)quinazoline-2,4-dione,melting point >300° C.

6-E-(4-methyl-1-pentenyl)!-3-(carboxymethyl)-1-(N-methyl-3-indolylmethyl)quinazoline-2,4-dione,melting point >300° C.

6-E-(3,3-dimethyl-1-butenyl)!-3-(carboxymethyl)-1-(N-methyl-3-indolylmethyl)quinazoline-2,4-dione,melting point >300° C.

EXAMPLE 2

a)6-Iodo-3-(methoxycarbonylmethyl)-1-(N-methyl-3-indolyl)quinazoline-2,4-dione

12.0 g (33.3 mmol) of6-iodo-3-(methoxycarbonylmethyl)quinazoline-2,4-dione prepared as inExample 1a)-c) were suspended in 250 ml of DMF, and 0.2 g (66.6 mmol) ofpotassium carbonate was added. Then 16.5 g (49.9 mmol) of(N-methyl-3-indolylmethyl)trimethylammoniumiodide were added and themixture was refluxed for 6 h. The solvent was stripped off under reducedpressure and then water and ethyl acetate were added, and the productwas filtered off with suction and dried. 16.2 g (96.5%) of product wereobtained, R_(F) 0.6 (ethyl acetate/n-heptane, 2:1).

b)6-(1-Pentenyl)-3-(methoxycarbonylmethyl)-1-(N-methyl-3-indolyl)quinazoline-2,4-dione

2.5 g (4.97 mmol) of the product obtained in a) were introduced into 20ml of DMF, and 1.46 g (14.9 mmol) of potassium acetate, 1.74 g (24.8mmol) of 1-pentene, 1.6 g (4.97 mmol) of tetrabutylammonium bromide and28 mg of palladium(II) acetate were added. The mixture was stirred atroom temperature for 16 h. The solvent was stripped off under reducedpressure, and the residue was taken up in ethyl acetate and 10% strengthEDTA solution. The organic phase was washed once more with EDTAsolution, dried and concentrated. The dark oily residue waschromatographed on silica with ethyl acetate/n-heptane (1:5) as eluent.1.23 g (55.5%) of product were obtained, R_(F) 0.61 (ethylacetate/n-heptane, 1:1).

c)6-Pentyl-3-(methoxycarbonylmethyl)-1-(N-methyl-3-indolyl)quinazoline-2,4-dione

0.2 g (0.45 mmol) of the product obtained in b) was dissolved in ethylacetate. After addition of 0.1 g of palladium/active carbon (10% Pd),the mixture was stirred under a hydrogen atmosphere for 6 h, duringwhich 15 ml of hydrogen were consumed. The catalyst was filtered offthrough silica gel, and the solution was washed with water, dried andconcentrated. 0.2 g of a pale yellow solid was obtained and wasprocessed further as the crude product.

d)6-Pentyl-3-(carboxymethyl)-1-(N-methyl-3-indolyl)quinazoline-2,4-dione

0.2 g (0.45 mmol) of the product obtained in c) was hydrolyzed withlithium hydroxide by the method of Example 1g. 0.11 g of product wasobtained, R_(F) 0.40 (dichloromethane, 20% methanol).

EXAMPLE 3

a)6-(2-Phenylethynyl)-3-(methoxycarbonylmethyl)-1-(N-methyl-3-indolyl)quinazoline-2,4-dione

3.5 g (6.95 mmol) of6-iodo-3-(methoxycarbonylmethyl)-1-(N-methyl-3-indolyl)quinazoline-2,4-dionewere dissolved in 50 ml of ethyl acetate and 1.42 g (13.9 mmol) ofphenylacetylene, 0.49 g (0.65 mmol) of bis(triphenylphosphine)palladiumdichloride, 0.066 g of copper(I) iodide and 3.52 g (34.8 mmol) oftriethylamine were added. The reaction mixture was refluxed for 8 h.Then 50 ml of ethyl acetate were added. Washing with 10% strength EDTAsolution was followed by drying and concentration. The solid brownresidue was chromatographed on silica gel with n-heptane/ethyl acetate(4:1) as eluent. 2.76 g (86%) of product were obtained, RF 0.19(n-heptane/ethyl acetate, 2:1).

b)6-(2-Phenylethynyl)-3-(carboxymethyl)-1-(N-methyl-3-indolyl)quinazoline-2,4-dione

1.5 g (3.25 mmol) of the product obtained in a) were reacted with 0.12 g(5.0 mmol) of lithium hydroxide as in Example 1g). The product wasinsoluble in water and ethyl acetate and was filtered off with suctionand dried. 0.78 g (52%) of product was obtained, RF 0.72(n-heptane/ethyl acetate, 1:2), melting point >300° C.

The following were prepared in a similar way:

6-(1-pentynyl)-3-(carboxymethyl)-1-(N-methyl-3-indolyl)quinazoline-2,4-dione

The following can be prepared in a similar way:

6-(4-methyl-1-pentynyl)-3-(carboxymethyl)-1-(N-methyl-3-indolyl)quinazoline-2,4-dione

6-(4-methyl-1-pentynyl)-3-(2-carboxyethyl)-1-(N-methyl-3-indolyl)quinazoline-2,4-dione

6-(4-methyl-1-pentynyl)-3-(3-carboxypropyl)-1-(N-methyl-3-indolyl)quinazoline-2,4-dione

EXAMPLE 4

a) 3-(2-Methylpropoxy)-6-nitrobenzaldehyde

50 g (0.308 mol) of 3-hydroxy-6-nitrobenzaldehyde and 49.3 g (0.36 mol)of 2-methylpropyl bromide were dissolved in 300 ml of DMF, and 45.5 g(0.33 mol) of potassium carbonate were added. The mixture was refluxedfor 5 h. After cooling, the solvent was stripped off under reducedpressure, and the residue was dissolved in ethyl acetate and washed with10% strength sodium carbonate solution and brine. The residue afterdrying and concentration was chromatographed on silica gel withn-heptane/ethyl acetate (9:1). 55.2 g (80%) of orange crystals wereobtained, R_(F) 0.43 (n-heptane/ethyl acetate, 9:1).

b) 3-(2-Methylpropoxy)-6-nitrobenzoic acid

24.2 g (0.067 mol) of t-butylammoniumpermanganate dissolved in 100 ml ofpyridine were added dropwise to a cooled solution of 22.3 g (0.10 mol)of the product obtained in a) in 150 ml of pyridine at such a rate thatthe temperature did not rise above 20° C. The mixture was stirred atroom temperature for 16 h and then poured into an ice/hydrochloric acidmixture and decolorized with Na₂ S₂ O₃. The aqueous phase was extractedwith ethyl acetate, the organic phase was made alkaline with ammonia,and the product was extracted into the aqueous phase. Afteracidification of the aqueous phase, the product was again extracted withethyl acetate, dried and concentrated. The product was recrystallizedfrom dichloromethane. 12.0 g (75%) of 3-(2-methylpropoxy)-6-nitrobenzoicacid were obtained.

c) 3-(2-Methylpropoxy)-6-aminobenzoic acid

8.8 g (36.8 mmol) of the product obtained in b) were dissolved in 50 mlof glacial acetic acid and, after addition of 1.5 g of palladium/activecarbon (10%), the mixture was stirred under a hydrogen atmosphere for 5h. The catalyst was filtered off with suction through silica gel, andthe solvent was stripped off under reduced pressure. Water was added tothe residue, and the product was filtered off with suction, washed withether and dried. 6.87 g (89%) of solid were obtained, R_(F) 0.3(n-heptane/ethyl acetate, 1:1).

d) 3-(2-Methylpropoxy)isatoic anhydride

6.87 g (32.8 mmol) of the product obtained in c) were converted into theisatoic anhydride as in Example 1a). 6.51 g (84%) of crude product wereobtained and were further processed without purification.

e) 2-Amino-5-(2-methylpropoxy)-N-(2-methoxycarbonylethyl)benzamide

6.51 g (27.7 mmol) of the product obtained in d) were reacted withβ-alanine methyl ester hydrochloride as in Example 1b). The crudeproduct was chromatographed on silica gel with n-heptane/ethyl acetate(2:1). 5.21 g (64%) of product were obtained, R_(F) 0.2 (n-heptane/ethylacetate, 1:1).

f) 6-(2-Methylpropoxy)-3-(2-methoxycarbonylethyl)quinazoline-2,4-dione

5.21 g (17.7 mmol) of the product obtained in e) were reacted as inExample 1c). The crude product was chromatographed on silica gel withn-heptane/ethyl acetate (2:1). 3.28 g (58%) of product were obtained,R_(F) 0.28 (n-heptane/ethyl acetate, 1:1).

g)6-(2-Methylpropoxy)-3-(2-methoxycarbonylethyl)-1-(3-indolylmethyl)quinazoline-2,4-dione

0.35 g (8.76 mmol) of 60% sodium hydride was suspended in 40 ml of THF,and 2.34 g (7.3 mmol) of the product obtained in f) dissolved in 30 mlof DMF were added dropwise. After 30 min, 2.77 g (8.76 mmol) of freshlyprepared (3-indolylmethyl)trimethylammonium iodide dissolved in 40 ml ofDMF were added dropwise, and the mixture was stirred at room temperaturefor 18 h. The residue after the solvent had been stripped off underreduced pressure was taken up in ethyl acetate and washed with 5%strength citric acid solution and water. After drying and concentration,the crude product was chromatographed on silica gel with n-heptane/ethylacetate (2:1). 1.29 g (39%) of product were obtained, R_(F) 0.52(n-heptane/ethyl acetate, 1:1).

h)6-(2-Methylpropoxy)-3-(2-carboxyethyl)-1-(3-indolylmethyl)quinazoline-2,4-dione

1.23 g (2.7 mmol) of the product obtained in g) were hydrolyzed withlithium hydroxide by the method of Example 1f). The crude product waspurified by HPLC with acetonitrile/water. 0.56 g (48%) of product wasobtained, melting point 199°-201° C.

The following were prepared in a similar way:

6-(2-methylpropoxy)-3-(3-indolylethyl)-1-(carboxymethyl)quinazoline-2,4-dione

6-(2-methylpropoxy)-3-(3-indolylethyl)-1-(2-carboxyethyl)quinazoline-2,4-dione

6-(2-methylpropoxy)-3-(3-indolylethyl)-1-(3-carboxypropyl)quinazoline-2,4-dione

6-(3-methylbutoxy)-3-(2-carboxyethyl)-1-(3-indolylmethyl)quinazoline-2,4-dione.

The following can be prepared as in Examples 1-3 from the corresponding4- and 6-iodoisatoic anhydrides:

5-(1-pentenyl)-3-(2-carboxyethyl)-1-(N-methyl-3-indolylmethyl)quinazoline-2,4-dione

5-pentyl-3-(2-carboxyethyl)-1-(N-methyl-3-indolylmethyl)quinazoline-2,4-dione

5-(4-methyl-1-pentenyl)-3-(2-carboxyethyl)-1-(N-methyl-3-indolylmethyl)quinazoline-2,4-dione

5-(4-methylpentyl)-3-(2-carboxyethyl)-1-(N-methyl-3-indolylmethyl)quinazoline-2,4-dione

5-(3-methyl-1-butenyl)-3-(2-carboxyethyl)-1-(N-methyl-3-indolylmethyl)quinazoline-2,4-dione

5-(3-methylbutyl)-3-(2-carboxyethyl)-1-(N-methyl-3-indolylmethyl)quinazoline-2,4-dione

7-(1-pentenyl)-3-(2-carboxyethyl)-1-(N-methyl-3-indolylmethyl)quinazoline-2,4-dione

7-(1-pentyl)-3-(2-carboxyethyl)-1-(N-methyl-3-indolylmethyl)quinazoline-2,4-dione

7-(4-methyl-1-pentenyl)-3-(2-carboxyethyl)-1-(N-methyl-3-indolylmethyl)quinazoline-2,4-dione

7-(3-methyl-1-butenyl)-3-(2-carboxyethyl)-1-(N-methyl-3-indolylmethyl)quinazoline-2,4-dione

7-(3-methylbutyl)-3-(2-carboxyethyl)-1-(N-methyl-3-indolylmethyl)quinazoline-2,4-dione

EXAMPLE 5

a) 3-Iodo-6-(bromoacetylamino)-N-(2-methoxycarbonylethyl)benzamide

21.0 g (60.5 mmol) of 3-iodo-6-amino-N-(2-methoxycarbonylethyl)benzamideprepared as in Example 1b) were dissolved in 270 ml of dichloromethane,and 12.5 ml (90.7 mmol) of triethylamine were added. At -30° C., 7.8 ml(90.7 mmol) of bromoacetyl bromide in 80 ml of dichloromethane wereadded dropwise. The mixture was stirred at room temperature for 2 h andthen washed with 10% strength citric acid solution and sodiumbicarbonate solution, dried and concentrated. The product wasrecrystallized from dichloromethane, resulting in 26.1 g (92%) of paleyellow crystals.

b) 7-Iodo-4-(2-methoxycarbonylethyl)-1,4-benzodiazepine-3,5-dione

7.5 g (16 mmol) of the product obtained in a) were suspended in 100 mlof methanol and slowly added dropwise to a solution of 32 mmol of sodiummethanolate in 400 ml of methanol. The mixture was stirred for 20 h andthen poured into phosphate buffer (pH 7), the methanol was stripped offunder reduced pressure, the aqueous phase was extracted with ethylacetate, and the organic phase was dried and concentrated. The crudeproduct was chromatographed on silica gel with dichloromethane (+3%methanol). 2.6 g (41%) of product were obtained, R_(F) 0.49(dichloromethane, 7% methanol).

c)7-Iodo-1-(N-methyl-3-indolylmethyl)-4-(2-methoxycarbonylethyl)-1,4-benzodiazepine-3,5-dione

2.6 g (6.6 mmol) of the product obtained in b) were dissolved in 100 mlof DMF, 1.82 g (1.32 mmol) of potassium carbonate were added and then3.27 g (9.9 mmol) of (N-methyl-3-indolylmethyl)trimethylammonium iodidewere added. The mixture was refluxed for 6 h, and then the DMF wasstripped off under reduced pressure, and the residue was taken up inwater. The aqueous phase was extracted with ethyl acetate, and theorganic phase was dried and concentrated. The crude product waschromatographed on silica gel with ethyl acetate/n-heptane (1:1) aseluent. 2.1 g (59%) of product were obtained, R_(F) 0.57(dichloromethane, 7% methanol).

d)7-(1-Pentenyl)-1-(N-methyl-3-indolylmethyl)-4-(2-methoxycarbonylethyl)-1,4-benzodiazepine-3,5-dione

2.1 g (3.9 mmol) of the product obtained in c) and 1.63 g (11.7 mmol) ofpotassium carbonate were introduced into 40 ml of DMF, and 1.37 g (19.5mmol) of 1-pentene, 1.26 g (3.9 mmol) of tetrabutylammonium bromide and20 g of palladium(II) acetate were added. The mixture was stirred at 50°C. for 2 h and at room temperature for 16 h. The solvent was thenstripped off under reduced pressure, and the residue was chromatographedon silica gel with dichloromethane (+2% methanol). 1.55 g (3.3 mmol=84%)of product were obtained, R_(F) 0.65 (dichloromethane, 7% methanol).

e)7-(1-Pentenyl)-1-(N-methyl-3-indolylmethyl)-4-(2-carboxyethyl)-1,4-benzodiazepine-3,5-dione

0.55 g (1.16 mmol) of the product obtained in d) were hydrolyzed by themethod of Example 1g). The crude product was purified by HPLC (reversedphase material, acetonitrile/water). 0.22 g (0.48 mmol=41.4%) of productwas obtained.

The following can be prepared in a similar way:

7-(4-methyl-1-pentenyl)-1-(N-methyl-3-indolylmethyl)-4-(2-carboxyethyl)-1,4-benzodiazepine-3,5-dione

7-(3-methyl-1-butenyl)-1-(N-methyl-3-indolylmethyl)-4-(2-carboxyethyl)-1,4-benzodiazepine-3,5-dione

6-(4-methyl-1-pentenyl)-1-(N-methyl-3-indolylmethyl)-4-(2-carboxyethyl)-1,4-benzodiazepine-3,5-dione

6-(3-methyl-1-butenyl)-1-(N-methyl-3-indolylmethyl)-4-(2-carboxyethyl)-1,4-benzodiazepine-3,5-dione

8-(3-methyl-1-butenyl)-1-(N-methyl-3-indolylmethyl)-4-(2-carboxyethyl)-1,4-benzodiazepine-3,5-dione

EXAMPLE 6

a)7-pentyl-1-(N-methyl-3-indolylmethyl)-4-(2-methoxycarbonylethyl)-1,4-benzodiazepine-3,5-dione

1.0 g (2.1 mmol) of the substance of Example 5d) was hydrogenated as inExample 2b). Chromatography of silica gel with ethyl acetate/n-heptane(1:4) resulted in 0.63 g (63%) of product, R_(F) 0.53 (ethylacetate/n-heptane, 1:1).

b)7-Pentyl-1-(N-methyl-3-indolylmethyl)-4-(2-carboxyethyl)-1,4-benzodiazepine-3,5-dione

0.5 g (1.05 mmol) of the product obtained in a) was hydrolyzed as inExample 1g). The crude product was purified by HPLC (reversed phasematerial/acetonitrile/water). 0.20 g (0.43 mmol=41%) of product wasobtained.

The following can be prepared in a similar way:

7-(3-methylbutyl)-1-(N-methyl-3-indolylmethyl)-4-(2-carboxyethyl)-1,4-benzodiazepine-3,5-dione

7-(3-methylbutyl)-1-(N-methyl-3-indolylmethyl)-4-(3-carboxypropyl)-1,4-benzodiazepine-3,5-dione

6-(3-methylbutyl)-1-(N-methyl-3-indolylmethyl)-4-(2-carboxyethyl)-1,4-benzodiazepine-3,5-dione

8-(3-methylbutyl)-1-(N-methyl-3-indolylmethyl)-4-(2-carboxyethyl)-1,4-benzodiazepine-3,5-dione

We claim:
 1. A method for treating a mammal in need ofendothelin-antagonistic activity which comprises administering apharmaceutically acceptable amount of a composition as defined in claim4 of the formula I ##STR15## where 2of the radicals A, B, D and E are CHgroups and the 2 other radicals are CH groups or nitrogen atoms,Z¹ ishydrogen or halogen, C₁₋₆ -alkyl, or phenyl, phenyl-C₁₋₄ -alkyl,naphthyl or naphthyl-C₁₋₆ -alkyl which are unsubstituted or substitutedon the aromatic radical by C₁₋₆ -alkyl, C₁₋₆ -alkoxy, halogen, CF₃, NO₂or CN, or C₂₋₆ -alkenyl or C₂₋₆ -alkynyl, C₃₋₇ -cycloalkyl or one of thegroups --NHR⁴, --NR⁴ ₂, --OR⁴, --SO₂ NHR⁴, --SO₂ NR⁴ ₂, --COR⁴ or --CO₂R⁴ (with R⁴ meaning C₁₋₄ -alkyl, phenyl, phenyl-C₁₋₄ -alkyl, naphthyl ornaphthyl-C₁₋₄ -alkyl), Z² has one of the meanings indicated for Z¹ butis not hydrogen, or Z¹ and Z² together with B and D are also one of theradicals ##STR16## (where Z³ has one of the meanings indicated for Z¹,and M is a CH₂ or NH group), G is a direct linkage or the group CH--K(with K meaning hydrogen, C₁₋₆ -alkyl, or a phenyl, benzyl, naphthyl ornaphthylmethyl group which is unsubstituted or substituted in the arylmoiety by C₁₋₄ -alkyl, C₁₋₄ -alkoxy, halogen, CF₃, NO₂ or CN), K isalkyl or alkenyl with, in each case, up to 6 C atoms or the group##STR17## L is an alkylene, alkenylene or alkynylene group with, in eachcase, up to 6 C atoms or one of the groups ##STR18## (with R³ meaninghydrogen, C₁₋₄ -alkyl, benzyl or naphthylmethyl), ##STR19## (with Qmeaning C₁₋₆ -alkyl, aryl or CH₂ --R⁷ where R⁷ is phenyl or ##STR20##(R³ ═H, C₁₋₃ -alkyl, --CHO, --COO--C₁₋₃ -alkyl)) or ##STR21## R¹ is--CO₂ R⁴ (with R⁴ meaning hydrogen, C₁₋₄ -alkyl or benzyl), --CONR⁴ ₂,--OR⁴, --SR⁴, --SO₃ R⁴, --PO₃ R⁴ ₂ or tetrazolyl, and R² is ##STR22##(with R⁵ and R⁶ meaning hydrogen, C₁₋₄ -alkyl, --OR⁴ or --SR⁴) orhetaryl,and, where appropriate, the salts thereof with physiologicallytolerated acids, for the production of drugs for controlling diseasesinvolving an elevated endothelin level.
 2. A compound of the formula I##STR23## where 2of the radicals A, B, D and E are CH groups and the 2other radicals are CH groups or nitrogen atoms,Z¹ is hydrogen orhalogen, C₁₋₆ -alkyl, or phenyl, phenyl-C₁₋₄ -alkyl, naphthyl ornaphthyl-C₁₋₆ -alkyl which are unsubstituted or substituted on thearomatic radical by C₁₋₆ -alkyl, C₁₋₆ -alkoxy, halogen, CF₃, NO₂ or CN,or C₂₋₆ -alkenyl or C₂₋₆ -alkynyl, C₃₋₇ -cycloalkyl or one of thegroups, --OR⁴, --SO₂ NHR⁴, --SO₂ NR⁴ ₂, --COR⁴ or --CO₂ R⁴ (with R⁴meaning C₁₋₄ -alkyl, phenyl, phenyl-C₁₋₄ -alkyl, naphthyl ornaphthyl-C₁₋₄ -alkyl), Z² has one of the meanings indicated for Z¹ butis not hydrogen, or Z¹ and Z² together with B and D are also one of theradicals ##STR24## (where Z³ has one of the meanings indicated for Z¹,and M is a CH₂ or NH group), G is a direct linkage or the group CH--K(with K meaning hydrogen, C₁₋₆ -alkyl, or a phenyl, benzyl, naphthyl ornaphthylmethyl group which is unsubstituted or substituted in the arylmoiety by C₁₋₄ -alkyl, C₁₋₄ -alkoxy, halogen, CF₃, NO₂ or CN), K isalkyl or alkenyl with, in each case, up to 6 C atoms or the group##STR25## L is an alkylene, alkenylene or alkynylene group with, in eachcase, up to 6 C atoms or one of the groups ##STR26## (with R³ meaninghydrogen, C₁₋₄ -alkyl, benzyl or naphthylmethyl), ##STR27## (with Qmeaning C₁₋₆ -alkyl, aryl or CH₂ --R⁷ where R⁷ is phenyl or ##STR28##(R³ ═H, C₁₋₃ -alkyl, --CHO, --COO--C₁₋₃ -alkyl)) or ##STR29## R¹ is--CO₂ R⁴ (with R⁴ meaning hydrogen, C₁₋₄ -alkyl or benzyl), --CONR⁴ ₂,--OR⁴, --SR⁴, --SO₃ R⁴, --PO₃ R⁴ ₂ or tetrazolyl, and R² is ##STR30##(with R⁵ and R⁶ meaning hydrogen, C₁₋₄ -alkyl, --OR⁴ or --SR⁴) orhetaryl,and, where appropriate, the salts thereof with physiologicallytolerated acids.
 3. A compound of the formula I ##STR31## where 2of theradicals A, B, D and E are CH groups and the 2 other radicals are CHgroups or nitrogen atoms,Z¹ is hydrogen or halogen, C₁₋₆ -alkyl, orphenyl, phenyl-C₁₋₄ -alkyl, naphthyl or naphthyl-C₁₋₆ -alkyl which areunsubstituted or substituted on the aromatic radical by C₁₋₆ -alkyl,C₁₋₆ -alkoxy, halogen, CF₃, NO₂ or CN, or C₂₋₆ -alkenyl or C₂₋₆-alkynyl, C₃₋₇ -cycloalkyl or one of the groups --NHR⁴, --NR⁴ ₂, --OR⁴,--SO₂ NHR⁴, --SO₂ NR⁴ ₂, --COR⁴ or --CO₂ R⁴ (with R⁴ meaning C₁₋₄-alkyl, phenyl, phenyl-C₁₋₄ -alkyl, naphthyl or naphthyl-C₁₋₄ -alkyl),Z² has one of the meanings indicated for Z¹ but is not hydrogen, or Z¹and Z² together with B and D are also one of the radicals ##STR32##(where Z³ has one of the meanings indicated for Z¹, and M is a CH₂ or NHgroup), G is a direct linkage or the group CH--K (with K meaninghydrogen, C₁₋₆ -alkyl, or a phenyl, benzyl, naphthyl or naphthylmethylgroup which is unsubstituted or substituted in the aryl moiety by C₁₋₄-alkyl, C₁₋₄ -alkoxy, halogen, CF₃, NO₂ or CN), K is alkyl or alkenylwith, in each case, up to 6 C atoms or the group ##STR33## L is##STR34## R¹ is --CO₂ R⁴ (with R⁴ meaning hydrogen, C₁₋₄ -alkyl orbenzyl), --CONR⁴ ₂, --OR⁴, --SR⁴, --SO₃ R⁴, --PO₃ R⁴ ₂ or tetrazolyl,and R² is ##STR35## (with R⁵ and R⁶ meaning hydrogen, C₁₋₄ -alkyl, --OR⁴or --SR⁴) or ##STR36## (R³ ═H, C₁₋₃ -alkyl, --CHO, --COO--C₁₋₃ -alkyl),and, where appropriate, the salts thereof with physiologically toleratedacids.
 4. A pharmaceutical composition comprising a pharmaceuticallyacceptable excipient and a compound as defined in claim 2.